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Evaluation of some innovative process for extraction of functional extract from cashew apple waste (Anacardium

occidentale, L.)

Agroindústria Tropical

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WORKSHOP: VALORISATION OF TRADITIONAL PROCESSING OF INDIGENOUS AND

UNDERUTILISED FRUITS

Institute of Technology of Cambodia, Phnomenh, Cambodia January 14-16, 2013.

The workshop is funded under the project "International network on preserving safety and n utrition of indigenousfruits and their derivatives", by the Leverhulme

Trust, UK

Agroindústria Tropical

AUTORS :

Fernando Antonio PINTO DE ABREU (phD research work), Embrapa Brezil

Manuel Dornier, Claudie Dhuique, Fabrice Vaillant ,Max Reynes

presented

by Max Reynes

UMR (Research Unit)CIRAD-QUALISUD

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Agroindústria Tropical

INTEREST of the presentation For this seminary (1)

-PRESENTATION OF A EXTRACTION AND PURIFICATION PROCESS WAY FOR ANY OTHER FUNCIONAL MOLECULE -TECHNOLOGY PRESENTED HAVE

*TO BE COUPLED WITH OTHERS TECHNOLOGIES * TO BE EVALUATE FOR OTHER RAW MATERIALS

-

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Agroindústria Tropical

INTEREST of the presentation For this seminary (2)

THIS PROCESS IS ACTUALLY EVALUATED FOR VALORISATION OF OTHER INDOGENOUS FRUITS: -STEVIA ( stevioside& sweeetener) -ELAGITANIN ( fruits rich in polifenols) -?? - COLORANT MOLECULES WITH HIGHT POTENTIAL MARKET :

- RED. GREEN etc…

- HEALTHY MOLECULES AGAINST TRIGLICERIDES? ANTI-INFLAMATORY, ANTIGLYCEMIA? ANTI OBESITY

* TO BE EVALUATE FOR OTHER RAW MATERIALS

-

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Agroindústria Tropical

INTEREST of the presentation For this seminary (3)

-- COLORANT MOLECULES WITH HIGHT POTENTIAL MARKET / RED. GREEN

- HEALTHY MOLECULES AGAINST TRIGLICERIDES? ANTI-INFLAMATORY, ANTIGLYCEMIA? ANTI OBESITY -IN EACH COUNTRY? WHATS IS THE TRADITIONAL KNOW HOW?

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Lutein

Zeaxanthin

Indoles

Vitamin K

Potassium

Beta-Carotene

Vitamin A

Bioflavonoids

Vitamin C

Potassium

Vitamin C

Lycopene Anthocyanins

Anthocyanins

Vitamin C

Phenolics

Allium

Allicin

Green Yellow

Orange Red Blue

Purple White

Spinach

Cabbage

Lettuce

Broccoli

Carrots

Oranges

Bananas

Radishes

Tomatoes

Strawberries

Blueberries

Eggplant

Garlic

Onions

Ref: E.Rock-INRA France 2009

Main antioxydants in fruits (remember…)

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Scavenge or interact with free radicals

Quench singlet oxygen

CAROTENOIDS Absorb light Lipophilic

Bind to hydrophobic surfaces

Easily isomerized And oxidized

IMPORTANT PHYSICAL AND CHEMICAL PROPERTIES OF CAROTENOIDS

Kimura, M. & Rodriguez-Amaya, D.B. (1999), Brazil;

OBJECTIVES OF THE RESEARCH WORK PRESENTED

GENERAL

SPECIFICS

OPTIMISE PROCESS PARAMETERS FOR EXTRACTION CARTENOIDES COUPLING MACERATION AND ENZIMATIC TREATEMENT AND PRESS

EVALUATION OF TANGENTIAL MICROFILTRATION OF THE OBTAINED CAROTENOIDS EXTRACTS FOR CONCENTRATION AND DIAFILTRATION PROCESS FOR PURIFICATION

CARACTERIZE AND QUANTIFY CAROTENOIDS CONTENTS AN THE Cahew apple WASTE EXTRACTS

EVALUATE A INNOVATIVE PROCESS FOR EXTRACTION AND PURIFICATION OF CAROTENOIDS EXTRACTS FROM CAHEW APPLE WASTE FOR A COLORANT USE WITHOUT ANY SOLVANT WITHOUT HEAT

07

07

The molecules………….

PRINCIPAL CAROTENOIDS IN FOODS

b -Carotene

b -Cryptoxanthin OH

Lycopene

Lutein HO

OH

CAROTENOIDS CONSIDERED IMPORTANT TO HUMAN HEALTH

Zeaxanthin HO

OH

Lycopene

Lutein HO

OH

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The indogenous fruit……

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Nuts ( fruit)

cashew varieties : view of CCP 76 - EMBRAPA Cashew apple “fruit”

Cashew apple

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Area of cashew growth in the word

* In the world 3.350.929 tons of nuts :year

* Exemple of Brésil : 220.505 tonnes It is 2 millions of tons of cahew apple by year

Mainly in Viet Nam , India , etc… Nigeria , Côte d’Ivoire , Brésil

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The raw product

NOIX DE CAJOU CASHEW JUICE

CASHEW APPLE ON THE TREE APPLE PRESS

WASTE

Use of waste?

CASHEW APPLE

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DIFFICULT TO TAKE OFF

Juice extraction with press

Raw juice

Apple waste

Carotenoids EXTRACTION maceration

CONCENTRATION Microfiltration

PURIFICATION Diafiltration

EXTRAIT BRUT (Emulsion)

Extract concentrated

Purified extract

fibers

Cashew apple

The process

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07

The process………….

CAROTENOIDS EXTRACTION

MACERATION ENZYMES ( pectinase / amylase DOSIS) TEMPERATURE TIME

PRESSURAGE RATIO EAU / FIBRES FORCE APPLIQUÉE VITESSE DE ROTATION

SCREW PRESS VUE 10

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Clarified

juice stable Enzymatic treatment

Pulpy juice

• free enzymes • linked in bioreactor

sterile permeat

rétentat

Membrane 0,2 mm

T < 35 °C

Pulpy juice

stabilised

With hight quality Pasteurization

Membrane technology and juice(ref.Vaillant)

Water… Enzymatic treatment

Pulpy juice

• free enzymes • linked in bioreactor

rétentat

Membrane 0,2 mm

T < 35 °C

Membrane technology and juice(ref.Vaillant)

For extract purification

CAROTENOID EXTRACT

∧

∨

∨ ∨ ∨ ∨

∧

∧

∨

∨

∨ ∧

∧

∧

∧

↵

↵

1

Perméat module 01

Perméat module 02

Perméat module 03

Perméat module 04

2

3

4

5 6

7

8

9

10

MICROFILTRATION PILOT ESSAY

MEMBRANE TO BE ADAPTED -Membranes monotubulaires Al2O3

-0.2 µm

LE MODE DE CONDUITE : 3 ETAPES *Concentration volume not variable *concentration volume variable *Diafiltration volume constant

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Tangential Microfiltration: various membranes

Hollow fibers (organic) 1 mm

Tubular multichannel (ceramic) 1 cm

∧

∨

∨

∨ ∨ ∨ ∨

∧

∧

∨

∨

∨ ∧

∧

Vmax

Vmin ∧

∧

extrait brut = perméat ; eau = 0

perméat

extrait brut

eau . V

∨

X

. V

. V

. V

. V

. V

CONCENTRATION using same VOLUME

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FRV = 1 + Vp

Vb

∧

∨

∨

∨ ∨ ∨ ∨

∧

∧

∨

∨

∨ ∧

∧

Vmax

Vmin ∧

∧

↵

↵

Perméat module 01

Perméat module 02

Perméat module 03

Perméat module 04

CONCENTRATION A VOLUME CONSTANT : QV (extrait brut) = QV (perméat) ; QV (eau) = 0

Extrait brut

∧

∨

∨

∨ ∨ ∨ ∨

∧

∧

∨

∨

∨ ∧

∧

Vmax

Vmin ∧

∧

extrait brut = 0 ; eau = 0

extrait brut

X X

eau

perméat

. V

. V

. V

. V

. V

CONCENTRATION using VOLUME VARIABLE

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0

20

40

60

80

100

120

140

160

180

200

220

240

260

1 3 5 7 9 11 13 15 17 19 21

0

20

40

60

80

100

120

140

160

180

200

220

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260

1 3 5 7 9 11 13 15 17 19 210

20

40

60

80

100

120

140

160

180

200

220

240

260

1 3 5 7 9 11 13 15 17 19 210

20

40

60

80

100

120

140

160

180

200

220

240

260

1 3 5 7 9 11 13 15 17 19 21

x

Facteur de Réduction Volumique - FRV

Den

sité

du

Flu

x d

e Pe

rméa

t (

L.h

-1.m

-1)

Concentration (same volume)

Concentration ( volume variable)

With enzymatic treatment

Without enzyme treatment

EVOLUTION FRV (PTM = 3,2 BAR)

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CONCENTRATION and PURIFICATION OF THE EXTRACTS

Microfiltration à FRV 1

Microfiltration à FRV 5 et 10

Microfiltration à FRV maximal

Purification par diafiltration

Optimisation extraction

Impact : enzyme Number of cycles Pressure transmembrane

Impact FRV

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Pressage CYCLE NUMBER

Impact on the quality extracted evaluated

Pressage CYCLE NUMBER

Pressage CYCLE NUMBER

Pressage CYCLE NUMBER

(HPLC-DAD)

10 20 30 40 50 60 min 0

10 20 30 40 50 60 min 0

10 20 30 40 50 60 min 0

0

50

100

150

200

mAU

0

50

100

150

200

mAU

0

50

100

150

200

mAU

cis - NEOXANTHINE

cis - VIOLAXANTHINE

450 nm

450 nm

400 nm

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Extraits

Fresh cashew apple

Extracts

CARACTERISATION CAROTENOIDES CONTENTS PROFILE

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The molecules extracted quality………….

CARACTERISATION DU PROFIL DES CAROTENOIDES

10 20 30 40 50 60 min 0

10 20 30 40 50 60 min 0

10 20 30 40 50 60 min 0

0

50

100

150

200

mAU

0

50

100

150

200

mAU

0

50

100

150

200

mAU

b

cis - NEOXANTHINE

cis - VIOLAXANTHINE

450 nm

450 nm

400 nm

β - CRYPTOXANTHINE

β - CAROTENE

LUTEINE MUTATOXANTHINE

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Extraits

Pomme de cajou fraîche

Extraits

CARACTERISATION CAROTENOIDES CONTENTS PROFILE

10 20 30 40 50 60 min 0

10 20 30 40 50 60 min 0

10 20 30 40 50 60 min 0

0

50

100

150

200

mAU

0

50

100

150

200

mAU

0

50

100

150

200

mAU

450 nm

450 nm

400 nm

cis-AUROXANTHINE

trans-AUROXANTHINE

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Extracts

Fresh cashew apple

extract

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PHYTOENE

PHYTOFLUENE

ζ-CAROTENE

β-ZEACAROTENE NEUROSPORENE α-ZEACAROTENE

RUBIXANTHINE γ-CAROTENE LYCOPENE δ-CAROTENE

β-CAROTENE β-CAROTENE – 5,6 –

EPOXYDE

β-CAROTENE – 5,6 , 5’, 6’ - DIEPOXYDE

β-CRYPTOXANTHINE

ZEAXANTHINE

ANTHERAXANTHINE

VIOLAXANTHINE

NEOXANTHINE

NEOCHROME

LYCOXANTHINE

LYCOPHYLLE

β-CRYPTOXANTHINE 5,6 EPOXIDE

CRYPTOFLAVINE

MUTATOXANTHINE

LUTEOXANTHINE

AUROXANTHINE

α-CAROTENE

α-CRYPTOXANTHINE OR ZEINOXANTHINE

LUTEINE

LUTEINE – 5,6 – EPOXIDE (TARAXANTHINE)

FLAVOXANTHINE OR CRYSANTHEMAXANTHINE

(1)

(1) (1) (1)

(2) (2)

(2) (2)

(3)

(3) (3) (3)

(3) (3)

(4)

(4)

(4)

(4)

(4)

(5)

(5)

(4)

(5)

(5)

(5)

(5) (5)

(2)

(1)

(1)

LUTEOCHROME

AUROCHROME

Carotenoides pathways

(1) : Déssaturation

(2) : cyclisation

(3) : Hydroxyilation

(4) : Époxidation

(5) : Réarrangement époxy-furanoxyde Source : RODRIGUEZ-AMAYA (2009) 21

* PROFILES BETWEEN INTIAL AND CONCENTRATED EXTRACTS IDENTICAL

* SOME CAROTENOIDES NOT IDENTICAL BETWEN CAHSEW APPLE AND EXTRACTS

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CARACTERISATION CAROTENOIDES CONTENTS PROFILE

EXTRACTION TECHNOLOGY CAN MODIFY MOLECULES EXTRACTED……

Factor of Concentration - Fc

Total Caroténoïdes content - Cc (mg.kg-1)

Raw carotenoid content - p (g.kg-1)

IMPACT of the PROCESS ON THE EXTRACT QUALITY

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11 caroténoïdes identified et quantified par HPLC – DAD - MS Auroxanthines cis et trans (40%) ; lutéine (20%) ; β-cryptoxanthine (17%)

3,8

Extract Caracterization

Liquid, yellow Easy to disolve in water Rich in caroténoïdes

CONCLUSIONS

Industrial use as natural colorant with hight value

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PERSPECTIVES (1)

PATENT Cirad/Embrapa :BR 10 2012 0097610) 26/04/2012 (research during 3 years; 1 PHD)

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Orientation for future research actions for Preserving functional activities: *technologies without temperature, free of solvant * screening of functional activities example with cashew: research actual running

But for all the other indogenous fruits?,,,

PERSPECTIVES (2)

EVALUATION SOCIO ENOMIC OF THE INDOGENOUS FRUITS AND EVALUATION OF THE ALREDAY TRADITIONAL KNOW HOW

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MAIN CONDITIONS TO START SUCH RESEARCH - specific laboratories with innovative equipments for extraction, purification with or not membrane: .microwave, resins, CO2 sp., etc - functionality analysis or chemical molecules ( for use as pesticides subsitute) or healthy products (screening of healthy actions)

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Max Reynes

Merry Christmas and Happy New year!!